Tongluo Huoxue formula reduces ligamentum flavum fibrosis in lumbar spinal stenosis.

OBJECTIVE

Lumbar spinal stenosis (LSS) caused by ligamentum flavum hypertrophy (LFH) is a common degenerative spinal condition leading to spinal cord and nerve root compression. This study aims to investigate the therapeutic effects and underlying mechanisms of Tongluo Huoxue Formula (TLHXF) in treating LSS by modulating the TGF-β/Smad signaling pathway.

METHODS

The active compounds of TLHXF were identified using HPLC-MS/MS (high-performance liquid chromatography-mass spectrometry). Network pharmacology was used to predict potential targets, followed by molecular docking to evaluate interactions between the active compounds and key proteins in the TGF-β/Smad signaling pathway. An LFH rat model was established, and ligamentum flavum (LF) thickness was measured using MRI. Histopathological examination, western blot (WB), polymerase chain reaction (PCR), immunohistochemistry (IHC), and enzyme-linked immunosorbent assay (ELISA) were performed to assess collagen deposition, inflammation, and fibrosis.

RESULTS

HPLC-MS/MS identified five active compounds in TLHXF. Network pharmacology and molecular docking confirmed that these compounds bind to key proteins, such as TGF-β1 and Smad2/3, thereby modulating fibrosis and inflammation. In vivo, TLHXF significantly reduced LF thickness and collagen deposition and inhibited the expression of fibrosis-related proteins (TGF-β1, Smad2/3, and type I collagen), as well as pro-inflammatory cytokines (iNOS, TNF-α, and IL-1β).

CONCLUSION

TLHXF shows strong anti-fibrotic and anti-inflammatory effects through the inhibition of the TGF-β/Smad signaling pathway, indicating its potential as a therapeutic agent for LSS caused by LFH.